Type
Text
Type
Dissertation
Advisor
Hadjiargyrou, Michael | Entcheva, Emilia | Simon, Sanford | Meng, Yizhi.
Date
2015-12-01
Keywords
Biomedical engineering
Department
Department of Biomedical Engineering.
Language
en_US
Source
This work is sponsored by the Stony Brook University Graduate School in compliance with the requirements for completion of degree.
Identifier
http://hdl.handle.net/11401/77002
Publisher
The Graduate School, Stony Brook University: Stony Brook, NY.
Format
application/pdf
Abstract
Bone tissue engineering provides an alternative to bone grafting. Our laboratory previously developed two different types of electrospun scaffolds for bone tissue engineering, including a DNA containing PLGA/PELA scaffold for delivering genes and a collagen/PLLA scaffold for delivering cells. For this dissertation research, we firstly fabricated a DNA containing PLGA/PELA electrospun scaffold using the same method as previously developed, with the intent of delivering osteogenic growth factor genes for bone regeneration. However, the scaffold shrunk severely after being submerged into aqueous environment and failed to achieve high transfection efficiency in vitro. Subsequently, we used the collagen/PLLA scaffold (originally developed for cell delivery) and modified our approach by surface immobilization of DNA/lipid complexes and showed superior transfection efficiency in Hela cells. Hence, BMP-2 gene delivery was performed using this collagen/PLLA scaffold with surface adsorbed transfection complexes on MC3T3 cells. Significant increase of mRNA level of BMP-2 expression was observed after 3 days of transfection and as were other preosteoblast differentiation markers at the mRNA level by day 7. However, the BMP2 protein expression was not detected and augmentation of mineralization was not observed in vitro probably due to the cytotoxicity of the transfection reagent. In contrast, in an in vivo setting, the BMP2 transfection complex immobilized collagen/PLLA scaffold successfully transfected surrounding tissue with both mRNA level and protein level expression. In addition, the BMP2 transfection complex loaded scaffold successfully induced ectopic mineralization in mouse skeletal muscle. These results indicate that the surface adsorption strategy is a promising approach to immobilize gene delivery vehicles onto electrospun scaffolds for bone tissue engineering applications. | 106 pages
Recommended Citation
Zhao, Xia, "BMP-2 Gene Delivery using Electrospun Collagen/PLLA Scaffold with Surface Adsorbed Transfection Complexes for Bone Formation" (2015). Stony Brook Theses and Dissertations Collection, 2006-2020 (closed to submissions). 2866.
https://commons.library.stonybrook.edu/stony-brook-theses-and-dissertations-collection/2866